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目的 研究前列腺癌细胞RNA转染树突状细胞(DCs)诱导的细胞毒T淋巴细胞(CTL)对前列腺癌细胞的特异性杀伤作用。方法联合应用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)诱导培养小鼠骨髓单个核细胞向DCs分化,脂质体法转染RNA,流式细胞术测定转染后树突状细胞共刺激分子表达的变化,MTT法测定DCs对T淋巴细胞的刺激能力。ELISA法检测IFN-γ的分泌水平。LDH法测定DCs诱导的CTL对前列腺癌细胞的杀伤效果。结果用GM-CSF和IL-4诱导培养小鼠骨髓来源的单核细胞,体外诱导培养6 d后获得大量DCs;RNA转染后,DCs表面共刺激分子CD40、CD80、CD86和MHC-Ⅱ分子表达提高(P<0.05),刺激T淋巴细胞增殖能力明显提高(P<0.01);前列腺癌RNA转染的DCs诱导CTL对前列腺癌细胞和肝癌细胞的杀伤作用有明显差异(P<0.01)。结论前列腺癌细胞RNA转染DCs后诱导的CTL对前列腺癌有特异性杀伤作用,为进一步抗肿瘤疫苗的实验研究奠定了基础。
Objective To study the specific cytotoxicity of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) transfected prostate cancer cells on prostate cancer cells. Methods Combined with GM-CSF and IL-4, the bone marrow mononuclear cells of mice were induced to differentiate into DCs. Lipofectamine was used to transfect RNA to the flow cytometry The expression of costimulatory molecules in dendritic cells after transfection was determined by cytometry. The ability of DCs to stimulate T lymphocytes was determined by MTT assay. ELISA method to detect the secretion of IFN-γ. LDH assay DCs-induced CTL cytotoxicity on prostate cancer cells. Results Bone marrow-derived mononuclear cells were induced by GM-CSF and IL-4, and a large number of DCs were obtained after cultured for 6 days in vitro. After RNA transfection, costimulatory molecules CD40, CD80, CD86 and MHC-Ⅱ (P <0.05), and the ability of stimulating T lymphocyte proliferation was significantly increased (P <0.01). The cytotoxicity of CTL induced by prostate cancer RNA transfected DCs on prostate cancer cells and hepatoma cells was significantly different (P <0.01). Conclusions CTLs induced by DCs transfected with prostate cancer cells have specific killing effect on prostate cancer, which lays the foundation for the further study of antitumor vaccine.